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重组类蜘蛛拖丝蛋白——表达与纯化

Recombinant Dragline Silk-Like Proteins-Expression and Purification.

作者信息

Gaines William A, Marcotte William R

机构信息

Clemson University.

出版信息

AATCC Rev. 2011 Mar;11(2):75-79.

Abstract

Spider dragline silk is a proteinaceous fiber with impressive physical characteristics making it attractive for use in advanced materials. The fiber is composed of two proteins (spidroins MaSp1 and MaSp2), each of which contains a large central repeat array flanked by non-repetitive N- and C-terminal domains. The repeat arrays appear to be largely responsible for the tensile properties of the fiber, suggesting that the N- and C-terminal domains may be involved in self-assembly. We recently isolated the MaSp1 and MaSp2 N-terminal domains from and have incorporated these into mini-silk genes for expression in transgenic systems. Current efforts involve the development of expression vectors that will allow purification using a removable affinity tag for scalable protein purification.

摘要

蜘蛛拖牵丝是一种具有令人瞩目的物理特性的蛋白质纤维,这使其在先进材料的应用中颇具吸引力。该纤维由两种蛋白质(蜘蛛丝蛋白MaSp1和MaSp2)组成,每种蛋白质都包含一个大型中央重复序列阵列,两侧是非重复的N端和C端结构域。重复序列阵列似乎在很大程度上决定了纤维的拉伸性能,这表明N端和C端结构域可能参与了自组装过程。我们最近从[来源]中分离出了MaSp1和MaSp2的N端结构域,并将它们整合到微型丝基因中,以便在转基因系统中表达。目前的工作包括开发表达载体,该载体将允许使用可去除的亲和标签进行纯化,以实现可扩展的蛋白质纯化。

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